Generation of a Biotin-Tagged Dual-Display Phage

Phage display is widely used in biomedical research. One of the great advantages of phage display is the specificity of the connection of a foreign peptide exposed outside the capsid to the intended target. Secondary detection systems, which are often laborious and costly, are required to identify a...

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Bibliographic Details
Published in:Cells (Basel, Switzerland) Switzerland), 2024-10, Vol.13 (20), p.1696
Main Authors: De Plano, Laura Maria, Oddo, Salvatore, Bikard, David, Caccamo, Antonella, Conoci, Sabrina
Format: Article
Language:English
Subjects:
Amino acids
Antibiotics
bacteria
bacteriophage
Bacteriophage M13 - genetics
Biotin
Biotin - metabolism
Biotinylation
Cell Surface Display Techniques - methods
Cloning
Detectors
E coli
Genetic engineering
Genomes
Medical research
Peptide Library
Peptides
Peptides - genetics
Peptides - metabolism
Phage display
plasmid
Plasmids
Proteins
Streptavidin
Streptavidin - metabolism
Tetracycline
Tetracyclines
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Summary:Phage display is widely used in biomedical research. One of the great advantages of phage display is the specificity of the connection of a foreign peptide exposed outside the capsid to the intended target. Secondary detection systems, which are often laborious and costly, are required to identify and quantify the peptide/target interaction. In this study, we generated a novel dual-display phage to facilitate the detection and quantification of the peptide/target interaction. First, we generated a biotin-tagged phage by adding a small biotin-accepting peptide (sBT) to gene-3 of the M13K07 helper phage. Subsequently, we enhanced the M13K07 biotin-tagged phage by incorporating a selective peptide on gene-8, which is then exposed to the phage capsid. The exposed peptide acts as a probe to bind to a selective molecular target, whose interaction can be readily visualized thanks to the biotinylated phage. Our versatile dual-display phage exhibits high flexibility; by swapping the displayed peptide/probe, one can change the phage target while retaining the sBT gene in-frame with the pIII. We expect the generated biotin-tagged dual phages to be used as a multifunctional probe to couple with several streptavidin-biotin-based systems.
ISSN:2073-4409
2073-4409
DOI:10.3390/cells13201696